Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays

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• 作者：Meena Adhikari ; Ulrich Strych ; Jinsu Kim ; Heather Goux ; Sagar Dhamane ; Mohan-Vivekanandan Poongavanam ; Anna E. V. Hagstr枚m ; Katerina Kourentzi ; Jacinta C. Conrad ; Richard C. Willson
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：December 1, 2015
• 年：2015
• 卷：87
• 期：23
• 页码：11660-11665
• 全文大小：376K
• ISSN：1520-6882

文摘

We introduce the modification of bacteriophage particles with aptamers for use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin鈥揵iotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, 鈭?00 times lower than those of previously reported IgE assays.